High-Throughput Mapping of a Dynamic Signaling Network in Mammalian Cells

Author:

Barrios-Rodiles Miriam12345,Brown Kevin R.12345,Ozdamar Barish12345,Bose Rohit12345,Liu Zhong12345,Donovan Robert S.12345,Shinjo Fukiko12345,Liu Yongmei12345,Dembowy Joanna12345,Taylor Ian W.12345,Luga Valbona12345,Przulj Natasa12345,Robinson Mark12345,Suzuki Harukazu12345,Hayashizaki Yoshihide12345,Jurisica Igor12345,Wrana Jeffrey L.12345

Affiliation:

1. Program in Molecular Biology and Cancer, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada, M5G 1X5.

2. Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada, M5G 2M9.

3. Department of Medical Genetics and Microbiology, University of Toronto, Toronto, Ontario, Canada, M5S 1A8.

4. Department of Computer Science, University of Toronto, Toronto, Ontario, Canada, M5S 3H5.

5. Banting and Best Department of Medical Research, University of Toronto, Toronto, Ontario, Canada, M5G 1L6.

Abstract

Signaling pathways transmit information through protein interaction networks that are dynamically regulated by complex extracellular cues. We developed LUMIER (for luminescence-based mammalian interactome mapping), an automated high-throughput technology, to map protein-protein interaction networks systematically in mammalian cells and applied it to the transforming growth factor–β (TGFβ) pathway. Analysis using self-organizing maps and k -means clustering identified links of the TGFβ pathway to the p21-activated kinase (PAK) network, to the polarity complex, and to Occludin, a structural component of tight junctions. We show that Occludin regulates TGFβ type I receptor localization for efficient TGFβ-dependent dissolution of tight junctions during epithelial-to-mesenchymal transitions.

Publisher

American Association for the Advancement of Science (AAAS)

Subject

Multidisciplinary

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